Figure 6.

Nessy cells die faster in culture compared with wild-type cells. Nessy (Ly5^b) (closed circles) and C57BL/6 (Ly5^a) (open circles) splenocytes from 10-week-old mice mice were cultured separately (data not shown) or together (in a ratio of 3 : 2) in 96-well plates at 5 × 10⁶ cells/well with 1 μg/ml Concanavalin A. Cells were collected after (a) 0, (b) 3, (c) 20, (d) 26, (e) 45 or (f) 70 hr in culture and analysed by flow cytometry. Cells were labelled with antibodies against Ly5^a, Ly5^b, CD4 and CD8, and stained with 7AAD to detect dead cells. Graphs show %7AAD⁺ (dead) cells after gating on lymphocytes by size and granularity, on Ly5^a (C57BL/6) or Ly5^b (nessy) and then on CD4 or CD8. Data shown are from one of two experiments in which similar trends were observed. P values are indicated (* < 0·05, ** < 0·01, *** < 0·005).